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Find video protocols related to scientific articles indexed in Pubmed.
Sequence types 235, 111, and 132 predominate among multidrug-resistant pseudomonas aeruginosa clinical isolates in Croatia.
Antimicrob. Agents Chemother.
PUBLISHED: 07-28-2014
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A population analysis of 103 multidrug-resistant Pseudomonas aeruginosa isolates from Croatian hospitals was performed. Twelve sequence types (STs) were identified, with a predominance of international clones ST235 (serotype O11 [41%]), ST111 (serotype O12 [15%]), and ST132 (serotype O6 [11%]). Overexpression of the natural AmpC cephalosporinase was common (42%), but only a few ST235 or ST111 isolates produced VIM-1 or VIM-2 metallo-?-lactamases or PER-1 or GES-7 extended-spectrum ?-lactamases.
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Usefulness of Pneumotest-latex for direct serotyping of Streptococcus pneumoniae isolates in clinical samples.
J. Clin. Microbiol.
PUBLISHED: 04-23-2014
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This study evaluated the usefulness of the Pneumotest-Latex assay for serotyping Streptococcus pneumoniae isolates directly in clinical samples. With an agreement of 88.1% with a PCR-based reference method, this test can be a useful tool for this study purpose, especially in clinical laboratories that do not have access to nucleic acid amplification technologies.
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[Recommendations for prevention of community-acquired pneumonia with bacteremia as the leading form of invasive pneumococcal infections in the population of people over 50 years of age and risk groups above 19 years of age].
Pol. Merkur. Lekarski
PUBLISHED: 04-12-2014
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Invasive pneumococcal disease (IPD) is a main cause of mortality associated with pneumococcal infections. Although, IPD is regarding mainly small children and persons in the age > 65 years, the investigations showed that because of IPD exactly sick persons are burdened with the greatest mortality in the older age, rather than of children. The most frequent form of IPD is community acquired pneumonia (CAP) with the bacteremia. The presence of even a single additional risk factor is increasing the probability of the unfavorable descent of pneumococcal infection. The risk factors for IPD and/or pneumonia with bacteremia apart from the age are among others asthma (> 2 x), chronic obstructive pulmonary disease (COPD), sarcoidosis (4 x), idiopathic pulmonary fibrosis (5 x), bronchiectases (2 x), allergic alveolitis (1.9 x) and pneumoconiosis (2 x), type 1 diabetes (4.4 x), type 2 diabetes (1.2 x), autoimmune diseases (e.g. rheumatoid arthritis (4.2 to 14.9 x), kidney failure with the necessity to dialysis (12 x), immunosuppression, cardiovascular disease, alcoholism and cancers. Examinations show that the best method of IPD and CAP preventing are pneumococcal vaccinations. On the market for ages 23-valent polysaccharide vaccine (PPV23) is available covering close the 90% of IPD triggering stereotypes. Her role in preventing CAP is uncertain and the immunological answer after vaccination at older persons and after revaccination is weak. Widely discussed disadvantageous effects of growing old of the immunological system show on the benefit from applying the immunization inducing the immunological memory, i.e. of conjugated vaccines which are activating the T-dependent reply and are ensuring the readiness for the effective secondary response. Examinations so far conducted with conjugated 7-valent and 13-valent (PCV13) vaccines at persons in the age > 50 years are confirming these expectations. Also sick persons can take benefits from PCV13 applying back from so-called IPD risk groups in the age > 19 years. At these work research findings were described above PPV23 and PCV13 at adults and world recommendations of applying both vaccines in risk groups from 19 years up to the advanced years. Also Polish recommendations of optimum applying of these vaccines were presented. They are recommending applying PCV13 at first in them, while PPV23, if to her readings exist should be given to > or = 8 of weeks from PCV13. In persons > or = 19 years which earlier received 1 or should receive more PPV23 doses first PCV13 dose should be given after the year or later than the last PPV23 dose, and then again PPV23 > or = 8 of weeks from PCV13 and the second PPV23 dose not earlier than 5 years from last PPV23. If the PPV23 application seems to be justified, it is irrespective of the more previous state vaccination against pneumococci, PCV13 should be given to as first.
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Molecular analysis of vanA outbreak of Enterococcus faecium in two Warsaw hospitals: the importance of mobile genetic elements.
Biomed Res Int
PUBLISHED: 01-15-2014
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Vancomycin-resistant Enterococcus faecium represents a growing threat in hospital-acquired infections. Two outbreaks of this pathogen from neighboring Warsaw hospitals have been analyzed in this study. Pulsed-field gel electrophoresis (PFGE) of SmaI-digested DNA, multilocus VNTR analysis (MLVA), and multilocus sequence typing (MLST) revealed a clonal variability of isolates which belonged to three main lineages (17, 18, and 78) of nosocomial E. faecium. All isolates were multidrug resistant and carried several resistance, virulence, and plasmid-specific genes. Almost all isolates shared the same variant of Tn1546 transposon, characterized by the presence of insertion sequence ISEf1 and a point mutation in the vanA gene. In the majority of cases, this transposon was located on 50 kb or 100 kb pRUM-related plasmids, which lacked, however, the axe-txe toxin-antitoxin genes. 100 kb plasmid was easily transferred by conjugation and was found in various clonal backgrounds in both institutions, while 50 kb plasmid was not transferable and occurred solely in MT159/ST78 strains that disseminated clonally in one institution. Although molecular data indicated the spread of VRE between two institutions or a potential common source of this alert pathogen, epidemiological investigations did not reveal the possible route by which outbreak strains disseminated.
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Interventions to reduce colonisation and transmission of antimicrobial-resistant bacteria in intensive care units: an interrupted time series study and cluster randomised trial.
Lancet Infect Dis
PUBLISHED: 10-23-2013
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Intensive care units (ICUs) are high-risk areas for transmission of antimicrobial-resistant bacteria, but no controlled study has tested the effect of rapid screening and isolation of carriers on transmission in settings with best-standard precautions. We assessed interventions to reduce colonisation and transmission of antimicrobial-resistant bacteria in European ICUs.
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Abundance and diversity of plasmid-associated genes among clinical isolates of Enterococcus faecalis.
Plasmid
PUBLISHED: 07-18-2013
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Enterococcus faecalis, a normal compound of the human intestinal microbiome, plays an important role in hospital-acquired infections. Plasmids make a significant contribution to the acquisition of the novel traits such as antimicrobial resistance and virulence by this pathogen. The study investigated the plasmid content and the diversity of plasmid-associated genes in a group of 152 hospital isolates of E. faecalis. The majority of plasmids visualized by pulsed-field gel electrophoresis of S1 nuclease-digested DNA fell into the range of 50-100 kb. PCR-based screening allowed detection of genes of the rep1(pIP501), rep2(pRE25), rep4(pMBB1), rep6(pS86), rep7(pT181), rep8(pAM373), rep9(pAD1/pTEF2/pCF10), rep10(pIM13) and rep13(pC194) families in 29 different combinations. The par and ?-?-? plasmid stabilization systems were ubiquitous (45 isolates, 29.6% and 88 isolates, 57.9%, respectively), while the axe-txe system was not found. The asa1 gene homologues encoding aggregation substance characteristic for the pAD1 and related group of pheromone-responsive plasmids were present in 106 isolates. A variety of sequence variants, including novel ones, of genes associated with pheromone-responsive plasmids, such as rep8(pAM373), rep9(pAD1/pTEF2/pCF10), par, and asa1 were observed. In conclusion, there is a big and only partially characterized pool of diverse plasmids in clinical E. faecalis.
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A decade of invasive meningococcal disease surveillance in poland.
PLoS ONE
PUBLISHED: 01-01-2013
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Neisseria meningitidis is a leading etiologic agent of severe invasive disease. The objective of the study was to characterise invasive meningococcal disease (IMD) epidemiology in Poland during the last decade, based on laboratory confirmed cases.
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Clinical and economic burden of community-acquired pneumonia among adults in the Czech Republic, Hungary, Poland and Slovakia.
PLoS ONE
PUBLISHED: 01-01-2013
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We estimate and describe the incidence rates, mortality, and cost of CAP (community-acquired pneumonia), in both inpatient and outpatient settings, in the Czech Republic (CZ), Slovakia (SK), Poland (PL), and Hungary (HU). A retrospective analysis was conducted on administrative data from the health ministry and insurance reimbursement claims with a primary diagnosis of pneumonia in 2009 to determine hospitalization rates, costs, and mortality in adults ?50 years of age. Patient chart reviews were conducted to estimate the number of outpatient cases. Among all adults ?50 years, the incidence of hospitalized CAP per 100,000 person years was: 456.6 (CZ), 504.6 (SK), 363.9 (PL), and 845.3 (HU). The average fatality rate for all adults ?50 is 19.1%, and for each country; 21.7% (CZ), 20.9% (SK), 18.6% (PL), 17.8% (HU). Incidence, fatality, and likelihood of hospitalization increased with advancing age. Total healthcare costs of CAP in EUR was 12,579,543 (CZ); 9,160,774 (SK); 22,409,085 (PL); and 18,298,449 (HU); with hospitalization representing over 90% of the direct costs of treatment. The burden of CAP increases with advancing age in four CEE countries, with hospitalizations driving the costs of CAP upwards in the elderly population. Mortality rates are generally higher than reported in Western EU countries.
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Insight into antimicrobial susceptibility and population structure of contemporary human Enterococcus faecalis isolates from Europe.
J. Antimicrob. Chemother.
PUBLISHED: 12-29-2011
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To investigate antimicrobial susceptibility and clonal relatedness of Enterococcus faecalis human isolates recovered recently (2006-09) in six European countries.
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A new rapid and cost-effective method for detection of phages, ICEs and virulence factors encoded by Streptococcus pyogenes.
Pol. J. Microbiol.
PUBLISHED: 12-22-2011
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Streptococcus pyogenes (group A Streptococcus, GAS) is a human pathogen that causes diseases of various intensity, from mild strep throat to life threatening invasive infections and postinfectional sequelae. S. pyogenes encodes multiple, often phage encoded, virulence factors and their presence is related to severity of the disease. Acquisition of mobile genetic elements, carrying virulence factors, as phages or ICEs (integrative and cojugative elements) has been shown previously to promote selection of virulent clones. We designed the system of eight low volume multi- and one singleplex PCR reactions to detect genes encoding twenty virulence factors (spd3, sdc, sdaB, sdaD, speB, spyCEP, scpA, mac, sic, speL, K, M, C, I, A, H, G, J, smeZ and ssa) and twenty one phage and ICE integration sites described so far for S. pyogenes. Classification of strains based on the phage and virulence factors absence or presence, correlates with PFGE MLST and emm typing results. We developed a novel, fast and cost effective system that can be used to detect GAS virulence factors. Moreover, this system may become an alternative and effective system to differentiate between GAS strains.
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[Invasive pneumococcal disease in Poland in 2010].
Pol. Merkur. Lekarski
PUBLISHED: 09-23-2011
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Streptococcus pneumoniae is the main etiologic agent of community-acquired invasive infections, especially in extreme age groups. Recently, the emergence of pneumococcal conjugate vaccines (PCV) brought a possibility to reduce the number of pneumococcal infections. Their introduction requires a knowledge concerning epidemiology of infections, which in different part of the world differs and changes with time, and therefore must be under permanent surveillance.
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Molecular characteristics of KPC-producing Enterobacteriaceae at the early stage of their dissemination in Poland, 2008-2009.
Antimicrob. Agents Chemother.
PUBLISHED: 09-19-2011
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After the first report in May 2008, the National Reference Center for Susceptibility Testing confirmed 113 cases of infection or colonization by KPC-producing members of the family Enterobacteriaceae in Poland by the end of 2009. The vast majority of patients were found in 18 hospitals; three patients were diagnosed at outpatient clinics. Most of the institutions were in the Warsaw area, including three hospitals with the highest numbers of cases. When available, the data on previous hospitalizations often indicated that these hospitals were the probable acquisition sites; one patient arrived from New York. The group of 119 unique isolates consisted of Klebsiella pneumoniae (n = 114), followed by Klebsiella oxytoca (n = 3), and Escherichia coli (n = 2). The K. pneumoniae isolates were dominated by the clone sequence type 258 (ST258) (n = 111); others were ST11 and ST23. The ST258 group was heterogeneous, with 28 pulsed-field gel electrophoresis (PFGE) subtypes, ?25 plasmid profiles, and nine ?-lactamase patterns differing by KPC variants (KPC-2 mainly), and SHV-12, CTX-M-3, and TEM-1-like enzymes. Plasmids carrying bla(KPC) genes varied in size (~48 to 250 kb), structure, and conjugation potential. Transferable IncFII(K) plasmids of ~110 to 160 kb, probably pKpQIL or its derivatives, were observed in all K. pneumoniae clones and in K. oxytoca. Also prevalent were nontypeable pETKp50-like plasmids of ~50 kb, found in K. pneumoniae ST258 and E. coli isolates (ST93 and ST224). Two K. pneumoniae-E. coli pairs from single patients might represent the in vivo transfer of such plasmids. The striking diversity of KPC producers at the early stage of dissemination could result from several introductions of these bacteria into the country, their multidirectional evolution during clonal spread, and transfer of the plasmids.
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Staphylococcal cassette chromosome mec (Sccmec) classification and typing methods: an overview.
Pol. J. Microbiol.
PUBLISHED: 09-13-2011
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Meticillin-resistant Staphylococcus aureus (MRSA) is one of the main causes of hospital-acquired infections, but since late 1990s also the community-acquired. For better understanding of the S.aureus epidemiology there is an urgent need for creation of new typing method for SCCmec element. The molecular typing of MRSA for epidemiological purposes is investigated by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), spa typing and the SCCmec type assignment. In last few years not only new type of SCCmec (VI to XI) have been identified, but also additional subtypes (i.e. IVg-j) and different variants of already existed one (i.e. 5C2&5 and 2B2&5) were discovered. The aim of this review is to briefly summarize current knowledge about SCCmec classification and to discuss advantages and disadvantages of selected SCCmec typing methods.
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Efficiency of the Cepheid Xpert vanA/vanB assay for screening of colonization with vancomycin-resistant enterococci during hospital outbreak.
Antonie Van Leeuwenhoek
PUBLISHED: 08-25-2011
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This study aimed to assess the efficiency of the Cepheid Xpert vanA/vanB test for detecting vancomycin-resistant enterococci (VRE) colonization during a VanA Enterococcus faecium outbreak and to compare the Cepheid Xpert vanA/vanB (Cepheid, Sunnyvale, USA) test to a culture method with chromogenic medium chromID VRE agar (bioMérieux). The Cepheid Xpert vanA/vanB assay showed sensitivity 61.5%, specificity 79.2%, positive predictive value 61.5% and negative predictive value 79.2%. The results obtained in this study demonstrate that a positive result in the Cepheid Xpert vanA/vanB test for vanA enables the rapid (less than 1 h) presumptive, prior to culture, recognition of patients colonized with VRE. However, the Cepheid Xpert vanA/vanB assay cannot be the only test used to screen patients during an ongoing VRE outbreak, because additional culturing of all samples negative for both vanA and vanB or positive for vanB should be performed in order to confirm the carrier status of the patient.
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Multilocus variable number tandem repeat analysis (MLVA) of Streptococcus pyogenes.
J. Microbiol. Methods
PUBLISHED: 07-12-2011
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We tested 21 polymorphic loci encoded by the genome of Streptococcus pyogenes (group A Streptococcus, GAS). Seven of them were chosen for the MLVA scheme. The primer pairs, designed for selected loci, detect from few to several alleles, and the method has a Simpsons Index of diversity of 0.957. To test the overall performance of the method, multiplex PCR reactions were carried out for over 700 GAS strains. Using the method we were able to detect differences between highly clonal strains that share the same emm, MLST and PFGE types. The most diverse strains were M4, M2, M3 and M28. We developed a typing method that can be employed to differentiate between GAS strains. The method has high resolution and measures diversity of the GAS core chromosome, on the contrary to methods such as PFGE.
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[Antibiotic resistance--what we have to do now?].
Pol. Merkur. Lekarski
PUBLISHED: 06-17-2011
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Antibiotics introduced about 60 years ago as miracle drugs are getting less effications. This is a result of the emergence and dynamic dissemination of resistance due to the clonal spread of resistant strains and horizontal gene transfer. This has decreased therapeutic options and has reached an alarming level, a threat to public health and patient care. The WHO and ECDC call for immediate action aimed at the introduction of rationale therapy, and the enhancement of microbiological diagnostics and infection control programs. Lack of action in this field may lead to a post antibiotic era.
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Comparison of multilocus variable-number tandem-repeat analysis with multilocus sequence typing and pulsed-field gel electrophoresis for Enterococcus faecalis.
Pol. J. Microbiol.
PUBLISHED: 01-01-2011
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Enterococcus faecalis represents recently an important etiological agent of health care-associated infections (HAIs) and there is a need for evaluation and comparison of typing methods available for this microorganism. We tested multilocus VNTR (variable-number tandem repeats) analysis (MLVA) on a well-characterized collection of 153 clinical isolates of E. faecalis, corresponding to 52 multilocus sequence types and 67 pulsed-field gel electrophoresis (PFGE) profiles. MLVA showed high discriminatory power, discerning 111 different types (diversity index equal 98.9%). The concordance MLVA/MLST and MLVA/PFGE was 0.95 and 0.74, respectively. High discriminatory power of MLVA indicates its utility for local epidemiology such as outbreak investigation, and for differentiation of clones defined by other methods.
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Gelatinase-associated phenotypes and genotypes among clinical isolates of Enterococcus faecalis in Poland.
Pol. J. Microbiol.
PUBLISHED: 01-01-2011
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Enterococcus faecalis is an important nosocomial pathogen causing serious invasive infections. One of the virulence factors of this pathogen, gelatinase GelE, is a protease whose gene expression is regulated by the Fsr quorum sensing system. In this study, we used a well-characterized collection of 153 clinical E. faecalis isolates to investigate the distribution of genes involved in gelatinase expression. Although 140 isolates (91% of the group) harbored the gelE gene, only 81 isolates (53%) produced active gelatinase. The gelatinase-negative phenotype was found in several unrelated clones, and appeared to be caused by various genetic events. Isolates of the hospital-adapted clonal complex 2 (CC2) and of CC40 were uniformly gelatinase-positive, while all the CC87 isolates contained the 23.9 kb deletion encompassing most of the fsr locus and were gelatinase-negative. No significant differences among isolates of different clinical origin and gelatinase activity or presence of the fsr genes were found with the exception of isolates from cerebrospinal fluid, which were more often gelatinase-positive than colonizing isolates.
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[European Antibiotic Awareness Day educational campaign--has it changed public attitudes to antibiotic use in Poland?].
Pol. Merkur. Lekarski
PUBLISHED: 11-24-2010
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Widespread inappropriate antibiotic use is a global concern for public health care. That is why national and international antibiotic resistance control strategies recommend education of health-care professionals and the public to promote prudent antibiotic use. This paper show the effect of the 2009 public antibiotic campaign in Poland on the knowledge and social attitudes towards use of antibiotics. The aim of this study was to assess the level of knowledge and behaviour models of Polish people regarding antibiotics, recommendations for their use as well as sources of information on antibiotics. At the same time the study was aimed at assessing whether actions undertaken during educational campaigns resulted in growing knowledge on the subject of antibiotic use or a change in attitudes towards this group of medicines.
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[BINet programme as a standard of modern health policy].
Med Wieku Rozwoj
PUBLISHED: 10-06-2010
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Infectious diseases--despite significant progress, both in their management and prevention--are becoming an increasingly serious problem, and their extent and character frequently constitute a threat for public health. In addition, a significant part of medical decisions regarding treatment and prevention of invasive bacterial infections is not made on the basis of the etiological agent of infection, which increases the chance of therapeutic failures, and makes the right decisions regarding prophylaxis difficult. In June 2008 the National Reference Centre for Bacterial Meningitis (Krajowy O?rodek Referencyjny ds. Diagnostyki Bakteryjnych Zakaze? O?rodkowego Uk?adu Nerwowego, KOROUN) started the BINet programme, which is a network monitoring invasive bacterial infections acquired outside the hospital. The programme aims to improve the diagnostic and thus therapeutic standards, in community acquired invasive bacterial infections in Poland. The BINet programme is based on the cooperation of clinicians, epidemiologists, microbiologists, and is headed by a team of the National Reference Centre for Bacterial Meningitis. The BINet programme enables quick and appropriate microbiological diagnosis of infections which is the basis of rational therapy and plays a key role in dealing with the infected patient. In addition, accurate epidemiological data allow to take quick and effective preventive measures at local and national level.
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Complex nature of enterococcal pheromone-responsive plasmids.
Pol. J. Microbiol.
PUBLISHED: 08-26-2010
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Pheromone-responsive plasmids constitute a unique group of approximately 20 plasmids identified, as yet, only among enterococcal species. Several of their representatives, e.g. pAD1, pCF10, pPD1 and pAM373 have been extensively studied. These plasmids possess a sophisticated conjugation mechanism based on response to sex pheromones--small peptides produced by plasmid-free recipient cells. Detailed analysis of regulation and function of the pheromone response process revealed its great complexity and dual role--in plasmid conjugation and modulation of enterococcal virulence. Among other functional modules identified in pheromone plasmids, the stabilization/partition systems play a crucial role in stable maintenance of the plasmid molecule in host bacteria. Among them, the par locus of pAD1 is one of the exceptional RNA addiction systems. Pheromone-responsive plasmids contribute also to enterococcal phenotype being an important vehicle of antibiotic resistance in this genus. Both types of acquired vancomycin resistance determinants, vanA and vanB, as well many other resistant phenotypes, were found to be located on these plasmids. They also encode two basic agents of enterococcal virulence, i.e. aggregation substance (AS) and cytolysin. AS participates in mating-pair formation during conjugation but can also facilitate the adherence ofenterococci to human tissues during infection. The second protein, cytolysin, displays hemolytic activity and helps to invade eukaryotic cells. There are still many aspects of the nature of pheromone plasmids that remain unclear and more detailed studies are needed to understand their uniqueness and complexity.
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The current status of invasive pneumococcal disease in Poland.
Vaccine
PUBLISHED: 07-23-2010
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The objectives of this study were to assess the incidence of invasive pneumococcal disease (IPD) in Poland (2006-2009), where mass vaccination had not been implemented, and to determine the serotype distribution and antimicrobial susceptibility of Streptococcus pneumoniae isolates. The IPD incidence rates were highest among children under 2 years of age (3.39/100,000 in 2009) and children 2-5 years old (2.44/100,000). The most common serotypes were 14, 3, 1, 4, 19F, 23F, 6B, and 12F (61.7% of all isolates). In children aged less than 5 years, isolates of serotypes 14, 6B, and 19F were most prevalent (52.7% of the IPD cases). The PCV7, PCV10, and PCV13 covered 43.3%, 54.8%, and 68.8% of all IPD cases, and 68.7%, 76.3%, and 86.3% of cases involving children under 5 years of age. Penicillin resistance was found in 21.3% of the isolates responsible for meningitis and in 1.2% of isolates responsible for other invasive infections. Introduction of antipneumococcal conjugated vaccines into the national immunisation programme would likely lead to a significant reduction of IPD-associated morbidity among Polish children in particular, as well as in the population as a whole, especially in cases involving pneumococci with a decreased susceptibility to antibiotics.
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[First isolation of metallo-beta-lactamase producing Klebsiella pneumoniae strain in Poland].
Anestezjol Intens Ter
PUBLISHED: 07-09-2010
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Metallo-beta-lactamases (MBL) are the enzymes that are able to hydrolyse almost the full range of beta-lactame antibiotics--penicillins, cephalosporins and carbapenems. The latter are the drugs of choice for treatment of serious infections caused by Enterobacteriaceae strains, which produce extended-spectrum-beta-lactamases. The presence of MBL-producing strains markedly decreases the therapeutic possibilities in severe, life-threatening infections.
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Multicenter study for defining the breakpoint for rifampin resistance in Neisseria meningitidis by rpoB sequencing.
Antimicrob. Agents Chemother.
PUBLISHED: 07-06-2010
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Identification of clinical isolates of Neisseria meningitidis that are resistant to rifampin is important to avoid prophylaxis failure in contacts of patients, but it is hindered by the absence of a breakpoint for resistance, despite many efforts toward standardization. We examined a large number (n = 392) of clinical meningococcal isolates, spanning 25 years (1984 to 2009), that were collected in 11 European countries, Argentina, and the Central African Republic. The collection comprises all clinical isolates with MICs of > or = 0.25 mg/liter (n = 161) received by the national reference laboratories for meningococci in the participating countries. Representative isolates displaying rifampin MICs of < 0.25 mg/liter were also examined (n = 231). Typing of isolates was performed, and a 660-bp DNA fragment of the rpoB gene was sequenced. Sequences differing by at least one nucleotide were defined as unique rpoB alleles. The geometric mean of the MICs was calculated for isolates displaying the same allele. The clinical isolates displaying rifampin MICs of > 1 mg/liter possessed rpoB alleles with nonsynonymous mutations at four critical amino acid residues, D542, H552, S548, and S557, that were absent in the alleles found in all isolates with MICs of < or = 1 mg/liter. Rifampin-susceptible isolates could be defined as those with MICs of < or = 1 mg/liter. The rpoB allele sequence and isolate data have been incorporated into the PubMLST Neisseria database (http://pubmlst.org/neisseria/). The rifampin-resistant isolates belonged to diverse genetic lineages and were associated with lower levels of bacteremia and inflammatory cytokines in mice. This biological cost may explain the lack of clonal expansion of these isolates.
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Population structure, virulence factors and resistance determinants of invasive, non-invasive and colonizing Streptococcus agalactiae in Poland.
J. Antimicrob. Chemother.
PUBLISHED: 06-27-2010
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To analyse Streptococcus agalactiae (group B Streptococcus; GBS) isolates collected in Poland from various human infections and carriage in respect of their clonality, distribution of virulence factors and antimicrobial resistance determinants, including the detection of transposons involved in the dissemination of antimicrobial resistance.
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Antimicrobial susceptibility of metallo-beta-lactamase positive and negative Klebsiella pneumoniae strains isolated from intensive care unit patients.
Pol. J. Microbiol.
PUBLISHED: 06-24-2010
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Carbapenem resistance in Gram-negative bacteria is a worldwide increasing and one of the most disturbing problems, given these antibiotics are drugs of choice in the treatment of infections caused by extended-spectrum-beta-lactamase producing strains. In this study the antibiotic susceptibility of metallo-beta-lactamase-positive and negative Klebsiella pneumoniae strains isolated from intensive care unit (ICU) patients was evaluated. The presence of genes encoding MBLs was determined with a commercial kit hyplex MBL ID (BAG HEALTH CARE). The MBL-producing isolates were the first K. pneumoniae isolates of this kind identified in Poland. It seems that methods for detecting MBLs in Enterobacteriaceae should be included in contemporary standards of microbiological diagnostics in the country.
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First report of Streptococcus pneumoniae serotype 6D isolates from invasive infections.
Vaccine
PUBLISHED: 04-26-2010
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We report the first invasive Streptococcus pneumoniae isolates of serotype 6D and the first occurrence of this serotype in Europe. Till now, the appearance of serotype 6D pneumococci in nasopharyngeal carriage has been speculated to be associated with a selective pressure from vaccination with the conjugated 7-valent antipneumococcal vaccine. Our observations indicate that this serotype is present also among unvaccinated individuals in the population where mass infant vaccination has not yet been introduced. Importantly, these strains were isolated from invasive infections, indicating the full virulence potential of pneumococci belonging to serotype 6D and its importance for future vaccine formulations.
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Characterization of transcription within sdr region of Staphylococcus aureus.
Antonie Van Leeuwenhoek
PUBLISHED: 04-20-2010
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Staphylococcus aureus is an opportunistic pathogen responsible for various infections in humans and animals. It causes localized and systemic infections, such as abscesses, impetigo, cellulitis, sepsis, endocarditis, bone infections, and meningitis. S. aureus virulence factors responsible for the initial contact with host cells (MSCRAMMs-microbial surface components recognizing adhesive matrix molecules) include three Sdr proteins. The presence of particular sdr genes is correlated with putative tissue specificity. The transcriptional organization of the sdr region remains unclear. We tested expression of the sdrC, sdrD, or sdrE genes in various in vitro conditions, as well as after contact with human blood. In this work, we present data suggesting a separation of the sdr region into three transcriptional units, based on their differential reactions to the environment. Differential reaction of the sdrD transcript to environmental conditions and blood suggests dissimilar functions of the sdr genes. SdrE has been previously proposed to play role in bone infections, whilst our results can indicate that sdrD plays a role in the interactions between the pathogen and human immune system, serum or specifically reacts to nutrients/other factors present in human blood.
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Expansion and evolution of the Streptococcus pneumoniae Spain9V-ST156 clonal complex in Poland.
Antimicrob. Agents Chemother.
PUBLISHED: 03-01-2010
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In this study, we analyzed 118 penicillin-nonsusceptible Streptococcus pneumoniae (PNSP) isolates (MICs, >or=0.12 microg/ml) recovered in Poland in 2003 to 2005 from patients with respiratory tract diseases and invasive infections. Seven different serotypes (14, 9V, 23F, 19F, 6B, 19A, and 6A, in order of descending frequency), seven alleles of the murM gene (murMA, murMB6, and the new murMB12 to -16 alleles), and 31 multilocus sequence types (STs) were observed. The vast majority of the PNSP isolates (90.7%) belonged to the international multiresistant clones, and among these, the Spain(9V)-ST156 clonal complex was the most prevalent (56 isolates) and was significantly overrepresented in invasive infections. The clone has been evolving rapidly, as demonstrated by the observed number of STs, the diversity in multiple-locus variable-number-tandem-repeat analysis (MLVA) types, and the polymorphism of pbp and pspA genes (coding for penicillin-binding proteins and the pneumococcal surface protein A, respectively). The presence and structure of the rlrA islet (encoding the pneumococcal pilus) were very well conserved. The Spain(9V)-ST156 clonal complex has been largely responsible for a decreasing susceptibility to penicillin among pneumococci in Poland in recent years, in spite of a relatively moderate antimicrobial use.
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The relBE2Spn toxin-antitoxin system of Streptococcus pneumoniae: role in antibiotic tolerance and functional conservation in clinical isolates.
PLoS ONE
PUBLISHED: 01-14-2010
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Type II (proteic) chromosomal toxin-antitoxin systems (TAS) are widespread in Bacteria and Archaea but their precise function is known only for a limited number of them. Out of the many TAS described, the relBE family is one of the most abundant, being present in the three first sequenced strains of Streptococcus pneumoniae (D39, TIGR4 and R6). To address the function of the pneumococcal relBE2Spn TAS in the bacterial physiology, we have compared the response of the R6-relBE2Spn wild type strain with that of an isogenic derivative, Delta relB2Spn under different stress conditions such as carbon and amino acid starvation and antibiotic exposure. Differences on viability between the wild type and mutant strains were found only when treatment directly impaired protein synthesis. As a criterion for the permanence of this locus in a variety of clinical strains, we checked whether the relBE2Spn locus was conserved in around 100 pneumococcal strains, including clinical isolates and strains with known genomes. All strains, although having various types of polymorphisms at the vicinity of the TA region, contained a functional relBE2Spn locus and the type of its structure correlated with the multilocus sequence type. Functionality of this TAS was maintained even in cases where severe rearrangements around the relBE2Spn region were found. We conclude that even though the relBE2Spn TAS is not essential for pneumococcus, it may provide additional advantages to the bacteria for colonization and/or infection.
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[European Antibiotic Awareness Day--why needed?].
Pol. Merkur. Lekarski
PUBLISHED: 11-26-2009
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Antibiotic resistance is a major European and global public health problem. There are a number of reasons for its emergence; however, the main reasons are the excessive and improper use of this group of antimicrobial drugs. Several recommendations on the prudent use of antibiotics have been developed by various national and intemational authorities. The main message is to use them more responsibly and rationally, since the very dynamic emergence and spread of resistant microorganisms, plus a lack of new antimicrobial drugs in the pipelines of pharmaceutical companies, may result in our inability to treat infections successfully in the near future. To strengthen the message on prudent use, an initiative of the ECDC resulted in a decision by the European Commission to establish the 18th of November as European Antibiotic Awareness Day every year.
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The effect of an antibiotic policy on the control of vancomycin-resistant enterococci outbreak and on the resistance patterns of bacteria isolated from the blood of patients in a hematology unit.
Pol. Arch. Med. Wewn.
PUBLISHED: 11-19-2009
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Antibiotic resistance has become one of the main medical problems worldwide. This is mainly due to an overuse and misuse of antibiotics.
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[Invasive pneumococcal disease in the Malopolska region of Poland, in the year 2002-2008. Is introduction of mass vaccination with conjugated pneumococcal vaccine justified?].
Med Wieku Rozwoj
PUBLISHED: 09-21-2009
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According to the WHO pneumococcal infections are the most common cause of morbidity and mortality in children.
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International clones of Klebsiella pneumoniae and Escherichia coli with extended-spectrum beta-lactamases in a Czech hospital.
J. Clin. Microbiol.
PUBLISHED: 08-26-2009
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A 2-month survey of extended-spectrum beta-lactamase (ESBL) producers was performed in a Czech hospital. Klebsiella pneumoniae produced SHV-2, -5, or -12, Escherichia coli produced CTX-M-9 or -15, and other species produced TEM-92 or -132. All K. pneumoniae and E. coli isolates belonged to sequence types (STs) or clonal complexes (CCs) spread across the world (K. pneumoniae clonal complex 11 [CC11], CC14, and sequence type 101 [ST101] and E. coli CC31, CC73, CC131, and CC405) and carried various plasmids (mainly with A/C- and FII-type replicons).
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[Peritonitis in the course of peritoneal dialisis caused by Haemophilus influenzae with BLNAR phenotype].
Pol. Merkur. Lekarski
PUBLISHED: 07-08-2009
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Most common bacterial species causing peritonitis in the course of peritoneal dialysis (PDP) are coagulase-negative staphylococci, Staphylococcus aureus and streptococci. Haemophilus influenzae is rarely associated with PDP. Hereby we present the first known case of APD-associated peritonitis caused by non-type able H. influenzae (NTHi) presenting the beta-lactamase negative, ampicillin-resistant (BLNAR) phenotype. An 18 year old boy who had been treated with the APD for 12 months due to SLE was admitted in good general condition with diagnosis of PDP. Standard diagnostic and therapeutical procedures were initiated. Dialysis fluid was turbid with cytosis of 435 WBC/ml. From dialysis fluid pure culture of Gram-negative coccobacillus was isolated. The isolate was identified as a BLNAR phenotype. The same bacterium was isolated from nasal swab. Blood cultures were negative. After evaluation of antimicrobial susceptibility the treatment was changed for the oral ciprofloxacin. The treatment was successful. Control tests 2 days later revealed cytosis of 15 WBC/mm3 and control cultures of peritoneal fluid were negative. After two weeks of treatment the patient was discharged in a good condition. Haemophilus influenzae is a bacterium frequently colonizing the nasopharyngeal cavity. A PCR-based method allowed to classify isolates as NTHi. Infection was probably of the respiratory origin as the isolates (from peritoneal fluid and nasal swab) were undistinguishable. There are only few reports describing this species as an ethiologic agent of peritonitis. This case prove that Haemophilus species should be taken into account as a possible aethiologic agent of PDP, especially in patients on immunosupression with carrier state of H. influenzae in the upper respiratory tract. This kind of microorganism requires specific conditions during its growing in vitro. Identification of its sensitivity to antibiotics is essential in order to detect strains of BLNAR phenotype, as it is a crucial part of an effective antibiotic therapy.
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Varicella vaccination in Europe - taking the practical approach.
BMC Med
PUBLISHED: 02-12-2009
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Varicella is a common viral disease affecting almost the entire birth cohort. Although usually self-limiting, some cases of varicella can be serious, with 2 to 6% of cases attending a general practice resulting in complications. The hospitalisation rate for varicella in Europe ranges from 1.3 to 4.5 per 100,000 population/year and up to 10.1% of hospitalised patients report permanent or possible permanent sequelae (for example, scarring or ataxia). However, in many countries the epidemiology of varicella remains largely unknown or incomplete. In countries where routine childhood vaccination against varicella has been implemented, it has had a positive effect on disease prevention and control. Furthermore, mathematical models indicate that this intervention strategy may provide economic benefits for the individual and society. Despite this evidence and recommendations for varicella vaccination by official bodies such as the World Health Organization, and scientific experts in the field, the majority of European countries (with the exception of Germany and Greece) have delayed decisions on implementation of routine childhood varicella vaccination, choosing instead to vaccinate high-risk groups or not to vaccinate at all. In this paper, members of the Working Against Varicella in Europe group consider the practicalities of introducing routine childhood varicella vaccination in Europe, discussing the benefits and challenges of different vaccination options (vaccination vs. no vaccination, routine vaccination of infants vs. vaccination of susceptible adolescents or adults, two doses vs. one dose of varicella vaccine, monovalent varicella vaccines vs. tetravalent measles, mumps, rubella and varicella vaccines, as well as the optimal interval between two doses of measles, mumps, rubella and varicella vaccines). Assessment of the epidemiology of varicella in Europe and evidence for the effectiveness of varicella vaccination provides support for routine childhood programmes in Europe. Although European countries are faced with challenges or uncertainties that may have delayed implementation of a childhood vaccination programme, many of these concerns remain hypothetical and with new opportunities offered by combined measles, mumps, rubella and varicella vaccines, reassessment may be timely.
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Automated peritoneal dialysis-associated peritonitis due to Haemophilus influenzae showing the BLNAR phenotype.
Int. J. Infect. Dis.
PUBLISHED: 01-10-2009
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A rare case of peritonitis due to beta-lactamase-negative, ampicillin-resistant Haemophilus influenzae is described in an 18-year-old male undergoing automated peritoneal dialysis. The infection was probably of respiratory origin. Two strains of H. influenzae cultured from peritoneal fluid and a nasal swab were indistinguishable by molecular methods. The patient was successfully treated with oral ciprofloxacin. The authors suggest that this species should be taken into account as the etiologic agent of peritoneal dialysis-associated peritonitis. Targeted antimicrobial therapy should be based on local antibiotic resistance patterns.
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New selective and differential chromogenic agar medium, chromID VRE, for screening vancomycin-resistant Enterococcus species.
J. Microbiol. Methods
PUBLISHED: 01-06-2009
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This study aimed to evaluate the usefulness of a novel differential culture medium, chromID VRE agar, for the isolation of VRE in a clinical laboratory. It was shown that ChromID VRE agar may be useful for rapid and selective isolation of VRE especially after inclusion of broth enrichment step.
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Multiple locus VNTR fingerprinting (MLVF) of Streptococcus pyogenes.
Virulence
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Streptococcus pyogenes (GAS) is a human pathogen that causes millions of infections worldwide. Comparison between GAS strains isolated in different laboratories all over the world is often difficult. Three usually used methods have either low resolution (emm typing), are expensive (MLST) or time- and labor-consuming (PFGE). Our laboratory recently developed new, inexpensive methods of GAS typing-VF (virulence factor profiling) and PP (phage profiling). To improve the typing scheme developed for GAS, we recently proposed a new typing method. Here we present detailed protocol for MLVF analysis.
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Susceptibility of Polish Bartonella henselae strains.
Pol. J. Microbiol.
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Due to the fastidious nature of B. henselae and the limited number of available isolates worldwide, there are few data on its in vitro susceptibility to antibiotics. We determined the minimal inhibitory concentrations (MIC) of ten antimicrobial agents against 11 feline isolates of B. henselae by Etest method. The lowest MICs were obtained for rifampicin < or = 0.002 mg/L. MICs of all isolates were < 0.016 mg/L for ampicilin, amoxicillin/clavulanic acid, tetracycline and ranged from 0.016 to 0.032 mg/L for azithromycin. The MICs for two tested fluoroquinolones: ciprofloxacin and levofloxacin ranged from 0.016 to 0.125 mg/L. The highest MICs were obtained for gentamicin ranging from 0.025 to 2.0 mg/L. Sulphonamide resistance genes sul 1, sul 2, sul 3 were not found in any of the tested isolates. Etest methodology seems to be a reliable method for determination of B. henselae susceptibility, however standardization is strongly desired.
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Detection of Streptococcus pyogenes virulence factors by multiplex PCR.
Virulence
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Streptococcus pyogenes (GAS) is a human pathogen that causes multiple infections worldwide. The pathogenic properties of GAS strains are often linked to the production of virulence factors such as toxins, proteases or DNases. Detection of virulence factors produced by GAS strains can be used to either determine pathogenic potential of the strain or as a rapid screening and typing method. We recently developed a method to detect simultaneously 20 GAS virulence factors (spd3, sdc, sdaB, sdaD, speB, spyCEP, scpA, mac, sic, speL, K, M, C, I, A, H, G, J, smeZ and ssa) in four low volume multiplex PCR reactions (Borek et al., 2011) and below we present a detailed protocol describing the method.
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Typing of Streptococcus pyogenes strains using the phage profiling method.
Virulence
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We recently developed a method that allows fast differentiation between Streptococcus pyogenes (GAS) strains. The method named phage profiling (PP) is based on a simple assumption that a regular PCR reaction with Taq polymerase and relatively short elongation time is not able to yield long DNA fragment, such as ~40-50 kb integrated prophage. Only fragments without any integrated DNA or short fragments inserted between integration sites can be efficiently amplified. We designed primers that anneal upstream and downstream prophage integration sites, so in simple PCR reaction we can test if any additional DNA is integrated into particular site. Profiling of integrated elements can be used as rapid, high resolution typing method, with the resolution as high as PFGE and is excellent predictor of PFGE type.
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[Invasive meningococcal disease in Opolskie Voivodeship between 2005-2010].
Pol. Merkur. Lekarski
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The article describes epidemiological situation concerning invasive meningococcal disease in Opolskie Voivodeship in comparison to the whole country in years 2005-2010, and activities undertaken related to the increased number of meningococcal infections. Owing to anti-epidemic and educational activities, distinct improvement of epidemiological situation in respect to invasive meningococcal disease in Opolskie Voivodeship was achieved. It required many of interdisciplinary activities and multi stage cooperation between Public Health Institutions. Only closed cooperation and exact realization of Meningococcal Disease Council recommendations and of the National Reference Centre for Bacterial Meningitis procedures permitted to control epidemic situation. Mass media played a significant role providing reliable and timely information and education campaign which allowed to reduce public anxiety in Opolskie Voivodeship.
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[Meningococcal infections].
Pol. Merkur. Lekarski
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Neisseria meningitidis is responsible for rapid-onset septicaemia and/or meningitis, the conditions that are referred as invasive meningococcal disease (IMD). Although cases of IMD usually appear only sporadically, they may also emerge in outbreaks and large epidemics. Due to epidemic potential, which Poland has been faced particularly in 2006-2007 and possible fulminant course of infection, many countries have established national meningococcal reference centre. Their activity focus on monitoring of IMD epidemiological situation, based on laboratory confirmed cases and possessed knowledge leads to optimal therapeutic and prophylactic decisions. In Poland such function is assigned to the National Reference Centre for Bacterial Meningitis (NRCBM).
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Nosocomial outbreak of Streptococcus pneumoniae Spain9V?ST156?14 clone in a pulmonary diseases ward.
Pol. Arch. Med. Wewn.
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 Streptococcus pneumoniae (S. pneumoniae) is one of the most common bacterial pathogens in community?acquired pneumonia. However, nosocomial pneumococcal infections are more and more widely observed.
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High genetic diversity among community-associated Staphylococcus aureus in Europe: results from a multicenter study.
PLoS ONE
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Several studies have addressed the epidemiology of community-associated Staphylococcus aureus (CA-SA) in Europe; nonetheless, a comprehensive perspective remains unclear. In this study, we aimed to describe the population structure of CA-SA and to shed light on the origin of methicillin-resistant S. aureus (MRSA) in this continent.
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